论文部分内容阅读
构建共表达HSV-1型gD和鼠源性IL-21DNA疫苗,探讨其是否能诱导动物免疫应答以及抵抗眼角膜HSV-1的感染。PCR扩增HSV-1gD和IL-21基因,分别定向插入载体pRSC中,构建DNA疫苗pRSC-gD-IL-21,对其进行鉴定后再分别以pRSC-gD-IL-21、pRSC-gD、pRSC免疫小鼠3次,间隔2周,末次免疫2周后检测血清抗体、IL-4及IFN-γ水平,特异性脾细胞增殖反应以及CTL、NK细胞杀伤活性。同时评价小鼠对病毒攻击角膜的免疫保护作用。经测序、酶切和RT-PCR鉴定后表明,pRSC-gD-IL-21中目的基因在细胞内成功转录表达。与对照鼠相比,该疫苗在鼠体内产生了较强的特异性抗体、IL-4及IFN-γ、特异性脾细胞增殖反应、CTL及NK细胞杀伤活性均增强,同时疫苗使小鼠产生了较强的针对眼角膜HSV-1感染免疫保护作用。结果显示,pRSC-gD-IL-21疫苗能诱导小鼠产生较强的免疫应答及针对HSV-1眼部感染的免疫保护作用。
To construct co-expression HSV-1 gD and murine IL-21 DNA vaccine and investigate whether it can induce animal immune response and resist the infection of corneal HSV-1. The recombinant plasmid pRSC-gD-IL-21 was transfected into pRSC vector pRSC-gD-IL-21 by PCR. The recombinant plasmids pRSC-gD-IL-21 and pRSC- The mice were immunized with pRSC three times at 2-week intervals, and the levels of serum antibody, IL-4 and IFN-γ, specific splenocyte proliferation and CTL and NK cell cytotoxic activity were measured 2 weeks after the last immunization. At the same time, the mice were evaluated for the protective immunity of the cornea against virus attack. Sequencing, restriction enzyme digestion and RT-PCR showed that the target gene in pRSC-gD-IL-21 was successfully transcribed in the cell. Compared with the control mice, the vaccine produced strong specific antibodies in mice. IL-4 and IFN-γ, specific splenocyte proliferation responses, cytotoxic activity of CTL and NK cells were enhanced, and the vaccine produced mice A strong protective effect against corneal HSV-1 infection. The results showed that pRSC-gD-IL-21 vaccine can induce mice to produce a strong immune response and protective immunity against HSV-1 eye infection.